# A Scalable 3D Human Liver Co-culture Platform for Hepatitis B Virus Infection

> **NIH NIH R21** · UNIVERSITY OF ILLINOIS AT CHICAGO · 2020 · $185,726

## Abstract

Abstract / Project Summary
Hepatitis B virus (HBV) infects ~240 million people chronically and can progress to cirrhosis and hepatocellular
carcinoma (HCC), the third leading cause of cancer mortality. Current drug therapies to treat HBV cannot
eliminate the persistent covalently closed circular (ccc)DNA, which thus requires lifetime drug therapy. Therefore,
the discovery of novel HBV drugs that can eliminate cccDNA is an active area of pharmaceutical drug
development. However, such efforts are hampered by the lack of physiologically-relevant model systems that
recapitulate critical features of the disease pathogenesis, such as interactions between hepatocytes and relevant
liver non-parenchymal cell types (NPCs), and the ability to evaluate patient-specific outcomes. Conducting tests
on the chimpanzee, the only animal model for HBV, is prohibitively expensive, severely restricted in the US and
Europe, and does not fully mimic human HBV pathogenesis. Thus, human liver culture platforms are the most
widely used model for HBV studies. While primary human hepatocytes (PHHs) are the gold standard for HBV
studies, they are a scarce resource that does not suffice for high-throughput drug screening and to elucidate the
genetic basis of HBV pathogenesis. On the other hand, induced pluripotent stem cell-derived human hepatocyte-
like cells (iHeps) are an important patient-specific source from an expandable precursor, thus mitigating many
of the limitations with PHHs. We and others have shown that 2D cultures of iHeps can be infected with HBV and
used to test drugs. However, absorbed extracellular matrix (ECM) proteins in 2D cultures do not allow adequate
elucidation of ECM reorganization/re-modeling during HBV-induced fibrosis progression. Furthermore, existing
platforms lack the relevant liver NPCs that interact with HBV-infected hepatocytes in vivo to modulate infection,
inflammation, and fibrosis. We have recently pioneered microscale 3D collagen microgels containing iHeps that
display adult-like liver functions, including drug metabolism enzyme activities, for several weeks in vitro when
co-cultured with primary liver sinusoidal endothelial cells (LSECs), herein referred to as ‘microtissues’. In this
proposal, we will test our novel hypothesis that iHep/LSEC 3D microtissues, which display adult-like liver
functions, can a) be infected chronically (weeks) with HBV with higher infection levels, spread, and amplification
than existing 2D/3D culture platforms (aim 1), and b) when augmented with hepatic stellate cells (HSCs) and
Kupffer cells (KCs), display inflammatory and fibrotic signatures correlative of clinical outcomes (aim 2). Our
proposal will yield a first-of-its-kind scalable 3D human liver platform containing iHeps and liver NPCs that
displays chronic HBV infection while retaining the ability to adequately metabolize drugs for screening. In the
future, our HBV platform can be used to study mechanisms underlying hepatocyte-NPC interactions toward...

## Key facts

- **NIH application ID:** 9951000
- **Project number:** 5R21AI147137-02
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT CHICAGO
- **Principal Investigator:** Salman R Khetani
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $185,726
- **Award type:** 5
- **Project period:** 2019-06-11 → 2022-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9951000

## Citation

> US National Institutes of Health, RePORTER application 9951000, A Scalable 3D Human Liver Co-culture Platform for Hepatitis B Virus Infection (5R21AI147137-02). Retrieved via AI Analytics 2026-05-27 from https://api.ai-analytics.org/grant/nih/9951000. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*